AKT-modified autologous intracoronary mesenchymal stem cells prevent remodeling and repair in swine infarcted myocardium  

作　　者：YU Yun-sheng SHEN Zhen-ya YE Wen-xue HUANG Hao-yue HUA Fei CHEN Yi-huan CHEN Ke LAO Wei-jie TAO Li 

机构地区：[1]Department of Cardiovascular Surgery, First Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215006, China

出　　处：《Chinese Medical Journal》2010年第13期1702-1708,共7页中华医学杂志（英文版）

摘　　要：Background Transplantation of adult bone marrow-derived mesenchymal stem cells （MSCs） has been proposed as a strategy for cardiac repair following myocardial damage. However cell transplantation strategies to replace lost myocardium are limited by the inability to deliver large numbers of cells that resist peritransplantation graft cell death. Accordingly, we set out to isolate and expand adult swine bone marrow-derived MSCs, and to engineer these cells to overexpress AKT1 （protein kinase B）, to test the hypothesis that AKTl-engineered MSCs are more resistant to apoptosis and can enhance cardiac repair after transplantation into the ischemic swine heart. Methods The CDS （regulation domain of AKT1） AKTI-cDNA fragment was amplified, and MSCs were transfected following synthesis with a pCDH1-AKT1 shuttling plasmid. Western blotting analysis and real-time reverse transcription-polymerase chain reaction （RT-PCR） was performed. Myocardial infarction （MI） models were constructed in Meishan pigs, and cardiac function was evaluated by magnetic resonance imaging （MRI） measurements and echocardiography 4 weeks later. All pigs were assigned to four groups： control （A）, DMEM （B）, MSC （C）, and AKT-transfected （D）. MSCs were transfected with the AKT1 gene, and autologous BrdU-labeled stem cells （1 × 10^7/5 ml） were injected into left anterior descending coronary atery （LAD） of the infarct heart in groups C and D. In group B, DMEM was injected using the same approach. In group A, there was no injection following LAD occlusion. After 4 weeks, cardiac function and regional perfusion measurements were repeated by MRI and echocardiography, and histological characteristics of the hearts were assessed. Connecxin-43 （CX-43）, BrdU, and von Willebrand factor （VWF） immunoreactivity was tested using enzyme linked immunosorbent assay （ELISA）. Vascular endothelial growth factor （VEGF）, transforming growth factor-β1 （TGF-β1） were analyzed at the same time. Results AKTI-cBackground Transplantation of adult bone marrow-derived mesenchymal stem cells （MSCs） has been proposed as a strategy for cardiac repair following myocardial damage. However cell transplantation strategies to replace lost myocardium are limited by the inability to deliver large numbers of cells that resist peritransplantation graft cell death. Accordingly, we set out to isolate and expand adult swine bone marrow-derived MSCs, and to engineer these cells to overexpress AKT1 （protein kinase B）, to test the hypothesis that AKTl-engineered MSCs are more resistant to apoptosis and can enhance cardiac repair after transplantation into the ischemic swine heart. Methods The CDS （regulation domain of AKT1） AKTI-cDNA fragment was amplified, and MSCs were transfected following synthesis with a pCDH1-AKT1 shuttling plasmid. Western blotting analysis and real-time reverse transcription-polymerase chain reaction （RT-PCR） was performed. Myocardial infarction （MI） models were constructed in Meishan pigs, and cardiac function was evaluated by magnetic resonance imaging （MRI） measurements and echocardiography 4 weeks later. All pigs were assigned to four groups： control （A）, DMEM （B）, MSC （C）, and AKT-transfected （D）. MSCs were transfected with the AKT1 gene, and autologous BrdU-labeled stem cells （1 × 10^7/5 ml） were injected into left anterior descending coronary atery （LAD） of the infarct heart in groups C and D. In group B, DMEM was injected using the same approach. In group A, there was no injection following LAD occlusion. After 4 weeks, cardiac function and regional perfusion measurements were repeated by MRI and echocardiography, and histological characteristics of the hearts were assessed. Connecxin-43 （CX-43）, BrdU, and von Willebrand factor （VWF） immunoreactivity was tested using enzyme linked immunosorbent assay （ELISA）. Vascular endothelial growth factor （VEGF）, transforming growth factor-β1 （TGF-β1） were analyzed at the same time. Results AKTI-c

关 键 词：gene therapy mesenchymal stem cell autologous implant myocardial infarction 

分 类 号：R686[医药卫生—骨科学]